title: Mass-scale high-throughput multiplex polymerase chain reaction for human platelet antigen single-nucleotide polymorphisms screening of apheresis platelet donors.
abstract: BACKGROUND
  Treatment with human platelet antigen (HPA)-matched platelets (PLTs) is the optimal therapy for bleeding secondary to neonatal alloimmune thrombocytopenia. Recent advances in high-throughput DNA-based blood group and PLT antigen genotyping have made it possible to screen plateletpheresis donors for potential HPA-matched PLT transfusion.
  
  
  METHODS
  This prospective study evaluated genomic DNA from plateletpheresis donors for single-nucleotide polymorphisms (SNPs) associated with HPA-1, -2, -3, -4, -5, and -15 to determine whether high-throughput multiplex genomic DNA PCR and oligonucleotide extension technology can be used for mass-scale PLT antigen genotyping. Genotyping using SNP technology was confirmed using sequence-specific polymerase chain reaction (SSP-PCR).
  
  
  RESULTS
  Of the 748 donors screened, 277 were found to be negative for antigens implicated in alloimmune thrombocytopenia. In addition, two donors were homozygous for HPA-1b/b and -2b/b, six donors for HPA-1b/b and -3b/b, one for HPA-2b/b and -3b/b, one for HPA-1b/b and -5b/b, 10 for HPA-1b/b and -15 b/b, four for HPA-5b/b and -15b/b, and one for HPA-2b/b and -15b/b. Retesting using SSP-PCR was conducted for 60 donors. Discrepant results occurred between SNP and SSP-PCR in less than 20% of samples for HPA-1b/1b/HPA-3b/3b, HPA-5b/5b, and HPA-15b/b.
  
  
  CONCLUSIONS
  High-throughput multiplex PCR SNP and confirmatory molecular genotyping are useful for mass-scale screening of apheresis PLT donors to provide antigen-negative genotypes. Refinements to mass-scale multiplex analysis technology would reduce further the confirmatory testing needed.
authors.affiliation: Department of Medicine, University of Toronto, Toronto, Ontario, Canada. shehatan@smh.toronto.on.ca
authors.forename: Nadine
authors.initials: N
authors.lastname: Shehata
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Denomme
authors.forename: Barbara
authors.initials: B
authors.lastname: Hannach
authors.forename: Nancy
authors.initials: N
authors.lastname: Banning
authors.forename: John
authors.initials: J
authors.lastname: Freedman
mesh_terms: D016824:Antigens, Human Platelet / Q000235:genetics*; D001782:Blood Donors; D046148:Donor Selection / Q000379:methods; D006801:Humans; D010983:Plateletpheresis; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011446:Prospective Studies; D054098:Thrombocytopenia, Neonatal Alloimmune / Q000628:therapy
chemical_list: D016824:Antigens, Human Platelet